Optimisation of protein solubility for NMR

		BioNMR > Sample preparation > Proteins
Optimisation of protein solubility for NMR

Webservers

NMR processing:

MDD

NMR assignment:

Backbone:

Side-chains:

NOEs:

Structure from NMR restraints:

Ab initio:

Fragment-based:

Rosetta-NMR (Robetta)

Template-based:

GeNMR

I-TASSER

Refinement:

Amber

Structure from chemical shifts:

Fragment-based:

Homology-based:

Torsion angles from chemical shifts:

Preditor

TALOS

Promega- Proline

Secondary structure from chemical shifts:

CSI (via RCI server)

TALOS

MICS caps, β-turns

d2D

PECAN

Flexibility from chemical shifts:

RCI

Interactions from chemical shifts:

HADDOCK

Chemical shifts re-referencing:

NMR model quality:

NOEs, other restraints:

Chemical shifts:

RDCs:

Pseudocontact shifts:

Protein geomtery:

SAVES2 or SAVES4

Quality Control Check

NMR spectrum prediction:

FANDAS

MestReS

V-NMR

Flexibility from structure:

Backbone S2

Methyl S2

B-factor

Molecular dynamics:

Gromacs

Amber

Antechamber

Chemical shifts prediction:

From structure:

Shiftx2

Sparta+

Camshift

CH3shift- Methyl

ArShift- Aromatic

ShiftS

Proshift

PPM

CheShift-2- Cα

From sequence:

Shifty

Camcoil

Poulsen_rc_CS

Disordered proteins:

MAXOCC

Format conversion & validation:

CCPN

From NMR-STAR 3.1

Validate NMR-STAR 3.1

NMR sample preparation:

Protein disorder:

DisMeta

Protein solubility:

Isotope labeling:

Solid-state NMR:

Thread Tools

Search this Thread

Rate Thread

Display Modes

08-14-2005, 07:53 PM

nmrlearner

Senior Member

Join Date: Jan 2005

Posts: 23,731

Points: 193,617, Level: 100

Level up: 0%, 0 Points needed

Activity: 50.7%

Last Achievements

Award-Showcase

NMR Credits: 0

NMR Points: 193,617

Downloads: 0

Uploads: 0

A straight-forward method of optimising protein solubility for NMR.

Howe PW.

Analytical Sciences, Syngenta, Jealott's Hill Research Centre, Bracknell, Berkshire, RG42 6EY, UK.

J Biomol NMR. 2004 Nov;30(3):283-6

Abstract:
Maximising solubility is a key step in applying solution-state NMR to proteins. The 'microbatch' crystallisation screening method can be adapted to screen for protein solubility. In this approach, drops of test solutions are placed under paraffin oil in 96-well screening plates. This requires very small amounts of protein, is easy to set up and is readily automatable.

Did you find this post helpful?

Similar Threads
Thread	Thread Starter	Forum	Replies	Last Post
Improving protein solubility & long-term stability Has anybody successfully used the following method? A simple method for improving protein solubility and long-term stability. Golovanov AP, Hautbergue GM, Wilson SA, Lian LY. Department of Biomolecular Sciences, University of Manchester Institute of Science and Technology, P. O. Box 88, Manchester M60 1QD, UK. a.golovanov@umist.ac.jp J Am Chem Soc. 2004 Jul 28;126(29):8933-9.	nmrlearner	Proteins	4	01-22-2014 07:04 PM
Supramolecular interactions between losartan and hydroxypropyl-?-CD: ESI mass-spectrometry, NMR techniques, phase solubility, isothermal titration calorimetry and anti-hypertensive studies. Supramolecular interactions between losartan and hydroxypropyl-?-CD: ESI mass-spectrometry, NMR techniques, phase solubility, isothermal titration calorimetry and anti-hypertensive studies. Supramolecular interactions between losartan and hydroxypropyl-?-CD: ESI mass-spectrometry, NMR techniques, phase solubility, isothermal titration calorimetry and anti-hypertensive studies. Int J Pharm. 2011 Feb 14;404(1-2):116-23 Authors: de Paula WX, Denadai AM, Santoro MM, Braga AN, Santos RA, Sinisterra RD In this work, low soluble supramolecular complex...	nmrlearner	Journal club	0	04-13-2011 11:57 PM
Overcoming the solubility limit with solubility-enhancement tags: successful applications in biomolecular NMR studies Overcoming the solubility limit with solubility-enhancement tags: successful applications in biomolecular NMR studies Abstract Although the rapid progress of NMR technology has significantly expanded the range of NMR-trackable systems, preparation of NMR-suitable samples that are highly soluble and stable remains a bottleneck for studies of many biological systems. The application of solubility-enhancement tags (SETs) has been highly effective in overcoming solubility and sample stability issues and has enabled structural studies of important biological systems previously deemed...	nmrlearner	Journal club	0	01-09-2011 12:46 PM
[NMR paper] A straight-forward method of optimising protein solubility for NMR. A straight-forward method of optimising protein solubility for NMR. Related Articles A straight-forward method of optimising protein solubility for NMR. J Biomol NMR. 2004 Nov;30(3):283-6 Authors: Howe PW Maximising solubility is a key step in applying solution-state NMR to proteins. The 'microbatch' crystallisation screening method can be adapted to screen for protein solubility. In this approach, drops of test solutions are placed under paraffin oil in 96-well screening plates. This requires very small amounts of protein, is easy to set up...	nmrlearner	Journal club	0	11-24-2010 10:03 PM
[NMR paper] NMR-based techniques in the hit identification and optimisation processes. NMR-based techniques in the hit identification and optimisation processes. Related Articles NMR-based techniques in the hit identification and optimisation processes. Expert Opin Ther Targets. 2004 Dec;8(6):597-611 Authors: Pellecchia M, Becattini B, Crowell KJ, Fattorusso R, Forino M, Fragai M, Jung D, Mustelin T, Tautz L In this review, the use of general NMR spectroscopy techniques to detect ligand binding and to monitor enzyme kinetics and inhibition, which appear particularly useful in hit identification and validation, is reiterated....	nmrlearner	Journal club	0	11-24-2010 10:03 PM
[NMR paper] A solubility-enhancement tag (SET) for NMR studies of poorly behaving proteins. A solubility-enhancement tag (SET) for NMR studies of poorly behaving proteins. Related Articles A solubility-enhancement tag (SET) for NMR studies of poorly behaving proteins. J Biomol NMR. 2001 May;20(1):11-4 Authors: Zhou P, Lugovskoy AA, Wagner G Protein-fusion constructs have been used with great success for enhancing expression of soluble recombinant protein and as tags for affinity purification. Unfortunately the most popular tags, such as GST and MBP, are large, which hinders direct NMR studies of the fusion proteins. Cleavage of the...	nmrlearner	Journal club	0	11-19-2010 08:32 PM

« Previous Thread | Next Thread »

Posting Rules
You may not post new threads You may not post replies You may not post attachments You may not edit your posts BB code is On Smilies are On [IMG] code is On HTML code is On Trackbacks are Off Pingbacks are Off Refbacks are Off